TGF-β-induced activation of conjunctival fibroblasts is modulated by FGF-2 and substratum stiffness

Abstract

Purpose This study aimed to investigate the effects of substratum stiffness on the sensitivity of human conjunctival fibroblasts to transforming growth factor (TGF)-β, and to explore the molecular mechanism of action. Methods Human conjunctival fibroblasts were cultured on collagen-coated plastic or silicone plates. The stiffness of the silicone plates was 0.2 or 64 kPa. Cells were treated by 2.5 ng/mL TGF-β2 with or without fibroblast growth factor (FGF)-2 (0–100 ng/mL) for 24 h or 48 h. The protein expression levels were determined by Western blot analysis. Cell proliferation was assessed using the WST-8 assay. Results FGF-2 suppressed the TGF-β-induced expression of α-smooth muscle actin (SMA) and collagen type I (Col I), but not fibronectin (FN). Both FGF-2 and TGF-β2 increased cell proliferation without an additive effect. The induction of α-SMA by TGF-β2 was decreased on the soft substratum, without any change in the expression level or subcellular location of Yes-associated protein/transcriptional coactivator with PDZ-binding motif (YAP/TAZ). FGF-2 suppressed TGF-β-induced α-SMA expression even on the soft substratum. Conclusions FGF-2 treatment and a soft substratum suppressed TGF-β-induced transdifferentiation of conjunctival fibroblasts into myofibroblasts. FGF-2 attenuated the TGF-β-induced expression of α-SMA, even on a soft substratum.

FGF-2 enhances fibrogenetic changes in TGF-β2 treated human conjunctival fibroblasts

“…For evaluating drug induced effects, 5 ng/mL solutions of TGF-β2, 10 ng/ml solutions of FGF-2, or both was added to 2D or 3D cultured Hcon F cells at Days 1 through 6. The dosages of the TGF-β2 and FGF-2 used in the present study were based on data reported in a previous study 25 , 26 .…”

“…In this area, Kay et al reported that FGF-2 stimulates TGF-β-mediated cell proliferation in rabbit subconjunctival fibroblasts 24 . On the contrary, Matsumura et al reported that FGF-2 induces the of the TGF-β-mediated trans-differentiation within human conjunctival fibroblast (HconF) cells 25 .…”

Suberoylanilide hydroxamic acid (SAHA) inhibits transforming growth factor-beta 2-induced increases in aqueous humor outflow resistance

“…The concentration of RNA samples was ascertained using a DS-11 NanoPad spectrophotometer (DeNovix, Wilmington, DE, USA). Reverse transcription and quantitative PCR were performed as previously described (63). Relative expression of the target mRNAs was compared to the control samples using the comparative threshold cycle method; glyceraldehyde-3-phosphate dehydrogenase was used as an endogenous control.…”

Pathophysiology of Pulmonary Fibrosis in the Context of COVID-19 and Implications for Treatment: A Narrative Review

“…The drug has been proven to inhibit TNF-alpha secretion in addition to other cytokines that cause the cytokine storm seen in SARS-CoV-2 patients [ 33 ]. Pirfenidone has also shown to decrease inflammatory response through inhibition of NLR family pyrin domain-containing (NLRP) 3, preventing the creation of an inflammasome complex that exacerbates COVID-induced inflammation [ 36 ]. In addition to its anti-inflammatory effects, pirfenidone also inhibits TGF- β -induced fibronectin synthesis as well as collagen 1 fibril formation [ 37 , 38 ].…”